摘要
目的:探讨尼美舒利对人食管癌Eca-109细胞生长和凋亡的影响及其作用机制。方法:应用MTT比色法检测尼美舒利对Eca-109细胞体外生长的抑制作用;流式细胞仪测定细胞周期和凋亡,琼脂糖凝胶电泳法进一步观察细胞凋亡;RT—PCR法检测COX-2、PPARγmRNA表达变化,Western blot检测PPARγ蛋白表达变化。结果:尼美舒利(50-400μmol/L)对Eca-109细胞生长有抑制作用,随浓度升高、时间延长作用增强,呈剂量-时间效应关系;可使Eca-109细胞G0/G1期比例增高,S期细胞减少,凋亡细胞增多并出现典型的凋亡细胞DNA ladder。此外,尼美舒利可下调COX-2表达并上调PPARγ表达。结论:尼美舒利能抑制Eca-109细胞的生长,其机制可能是通过抑制Eca-109细胞增殖、诱导细胞周期G0/G1期阻滞、细胞凋亡、下调COX-2表达、上调PPARγ表达而实现的。
Objective:To explore the effects and possible mechanism of nimesulide in inducing growth inhibition and apoptosis on human esophageal carcinoma Eca-109 cell line. Methods: Inhibition of Eca-109 cell growth induced by nimesulide at different concentrations was assayed by MTT method. Cell cycle and apoptosis were detected by flow cytometry, moreover. DNA ladder was separated by agarose gel electrophoresis. Alterations of cyclooxygenase-2 (COX-2), peroxisome proliferators-activated receptors γ(PPAR γ) mRNA expression were evaluated by reverse transcription polymerase chain reaction (RT-PCR). Western blot was used to analyze the expression of PPARγ protein expression. Results: Nimesulide 50-400 μmol/L inhibited the growth of Eca-109 cell line in a dose - and time -dependent manner. Nimesulide also increased the proportion of cells in G0/G1 phase,decreased the proportion of cells in S phase and induced apoptosis. Furthermore,nimesulide down-regulated the COX-2 mRNA expression and up-regulated the PPARγ expession. Conclusion: Nimesulide can inhibit the growth of Eca-109 cell line in vitro. The mechanism is probably associated with inhibition of cell proliferation, induction of G0/G1 arrest and apoptosis, down-regulation of the expression of COX-2 and up-regulation of the expression of PPART.
出处
《肿瘤》
CAS
CSCD
北大核心
2006年第3期244-248,共5页
Tumor
基金
河北省自然科学基金资助课题(编号:301354)